Separation Of Anticancer Compounds From The Euphorbia Lathyris And Its Pharmacological Research

Euphorbia lathyris has been known traditionally for its anticancer activity.However,for weakly polar compositions of Euphorbia lathyris,there is a lack of comprehensive chromatographic purification methods.Therefore,it is necessary to establish an efficient bioassay-guided off-line 2D HPLC for isolation and identification of anticancer compounds from the Euphorbia lathyris ethyl acetate extract.In present study,a Silica column coupled with an Amide column is employed to construct a novel nonaqueous 2D prep-HPLC separation system.Preparative separation in the first dimension was performed on a bare Silica column to simplify the sample into the fractions with good separation repeatability.Then,an Amide column was taken as the second dimension to realize the rapid preparation of monomers.Structures were established by 1D and 2D NMR spectroscopy.For compounds 2 absolute configuration was confirmed by X-ray single crystal diffraction spectroscopy.The compounds were evaluated for their tumor cell growth inhibitory activities on HEPG2 and A549 cells by real-time cell-analyzer.Different doses of Euphorbia factor L1 was given orally in mice to evaluate their acute toxicities.The in vivo anti-tumor experiments using Lewis lung cancer bearing mice,to investigate inhibitory effect on solid tumor and the life of the prolongation of lung cancer.Eventually,a total of seven compounds were isolated from the Euphorbia lathyris extract,namely Euphorbia factor L2(1),Euphorbia factor L1(2),Glyceryl monooleate(3),Euphorbia factor L8(4),Euphorbia factor L3(5),Euphorbia factor L9(6),and Esculetin(7).Compounds 2 and 6 showed significant anticancer activity against A549 and HEPG2 cells.Compounds 1,4 and 7 were inapparent against HEPG2 cells.However,the four compounds showed a certain degree bioactivity against A549 cells.Compound 5 exhibited a good inhibition of A549 cells growth activity than HEPG2 cells.Euphorbia factor L1 not cause obvious cytotoxicity in mice at a dose of 11.3 g/kg.In vivo,Euphorbia factor L1 inhibited Lewis mice transplant tumor,the inhibit rates being 18.22-57.53%.As mentioned above,the bioactivity-guided off-line 2D HPLC method is extremely useful for the separation of the weakly polar compounds and could lead to the discovery of more helpful natural anticancer compounds.